Dendritic Cells and Periodontal Disease

树突状细胞和牙周病

• 批准号: 8476210
• 负责人: DANA T GRAVES
• 金额: $ 38.4万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8476210
• 关键词: Address; Alveolar Bone Loss; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Bacteria; Binding; Biological Assay; Bone Resorption; Breeding; Calvaria; Cells; DNA; DNA Binding; Data; Dendritic Cells; Disease Progression; Down-Regulation; Event; Exhibits; Flow Cytometry; Gene Expression; Gene Targeting; Generations; Genetic Transcription; Goals; Histologic; ITGAX gene; Immune response; Immunization; In Vitro; Inflammatory; Inflammatory Response; Injection of therapeutic agent; Interferons; Interleukin-1; Interleukin-10; Interleukin-12; Interleukin-6; Link; Measurement; Measures; Mediating; Modeling; Monitor; Mus; Nuclear; Oral; Oral cavity; Osteoclasts; Pathogenesis; Periodontal Diseases; Periodontium; Play; Porphyromonas gingivalis; Process; Production; Promoter Regions; Regulatory Pathway; Reporting; Role; Scalp structure; Signal Transduction; Site; Spleen; TNF gene; Testing; Up-Regulation; base; bone; bone loss; chromatin immunoprecipitation; cytokine; in vivo; in vivo Model; interleukin-12 subunit p40; mutant; oral infection; osteoclastogenesis; pathogen; prevent; promoter; recombinase; research study; response; soft tissue; transcription factor

DESCRIPTION (provided by applicant): Dendritic cells (DC) may play a central role in the pathogenesis of periodontal disease by participating in events that link bacterial stimulation to periodontal bone loss. They may accomplish this by the production of factors that regulate the adaptive and innate immune responses. Furthermore, immature DCs have been reported to function as osteoclasts precursors. Preliminary Data examining DCs in vitro establish that the transcription factor FOXO1 is essential for LPS up-regulation of several cytokines, IL-1, TNF, IL-6, and IL-12 and down-regulation of the anti-inflammatory cytokine IL-10. Moreover, Akt plays an important role in this process by reducing FOXO1 nuclear localization and limiting inflammatory cytokine production in DC. Thus, we propose that FOXO1 is critical in upregulating an inflammatory response in DCs and that a hyper- inflammatory response is prevented by Akt. These findings serve as the basis for the current proposal in which we will test in vivo the hypothesis that the FOXO1-Akt axis regulates DC cytokine expression. Moreover, we will determine whether this regulatory pathway is essential for stimulating the adaptive immune response to the periodontal pathogen, P. gingivalis, and whether it plays a significant role in bacteria induced bone loss. To delete FOXO1 in DCs we will use the Cre-lox approach. We have already bred floxed FOXO1 mice with mice that express Cre recombinase under control of the CD11c promoter. DC isolated from the resulting experimental (CD11cCre+/FOXO1L/L) mice exhibit reduced cytokine expression stimulated by LPS compared to littermate control mice (CD11cCre-/FOXO1L/L). We will examine host-bacteria interactions in vivo by injecting P. gingivalis in the calvarial model. Because mice are naive to P. gingivalis we can examine the response when the adaptive immune response is not present and compare it to mice in which the adaptive immune response is activated by pre-immunization with P. gingivalis. By flow cytometry we will have a detailed analysis on the impact of FOXO1 deletion in generating a systemic and local adaptive immune response. The underlying calvarial bone will also be examined histologically to investigate the subsequent effect on osteoclastogenesis and bone resorption. The oral gavage model of applying P. gingivalis to the oral cavity will be studied in experimental (CD11cCre+/FOXO1L/L) and control mice (CD11cCre-/FOXO1L/L ) to determine whether FOXO1 deletion in DCs modulates the host response to P. gingivalis, osteoclastogenesis and periodontal disease progression. Experiments using the same two in vivo models will determine whether Akt is necessary to prevent a hyperinflammatory response in DC. These experiments will examine experimental (CD11cCre+/FOXO1L/L) and control (CD11cCre-/FOXO1L/L) mice. In Aim 3 the mechanisms by which the FOXO1-Akt axis regulates selected target gene expression (IL-1, TNF, IL-6, and IL-12) will be examined in vitro.

描述(申请人提供)：树突状细胞(DC)可能通过参与将细菌刺激与牙周骨丢失联系起来的事件，在牙周病的发病机制中发挥核心作用。它们可能通过产生调节适应性和先天免疫反应的因子来实现这一点。此外，据报道，未成熟的DC具有破骨细胞前体的功能。在体外检测DC的初步数据表明，转录因子FOXO1对于内毒素上调几种细胞因子IL-1、TNF、IL-6和IL-12以及下调抗炎细胞因子IL-10是必不可少的。此外，Akt通过减少FOXO1的核定位和限制DC中炎性细胞因子的产生而在这一过程中发挥重要作用。因此，我们认为FOXO1在上调DC的炎症反应中起关键作用，Akt可以预防高炎症反应。这些发现是当前提议的基础，我们将在体内测试FOXO1-Akt轴调节DC细胞因子表达的假设。此外，我们将确定这一调节途径是否对于刺激对牙周病原体P.gigivalis的适应性免疫反应是必不可少的，以及它是否在细菌诱导的骨丢失中发挥重要作用。要删除DC中的FOXO1，我们将使用CRE-LOX方法。我们已经用CD11c启动子控制下表达Cre重组酶的小鼠培育了FOXO1小鼠。从实验小鼠(CD11cCre+/FOXO1L/L)分离的DC在脂多糖刺激下的细胞因子表达低于同年龄对照组小鼠(CD11cCre-/FOXO1L/L)。我们将通过在颅骨模型中注射牙龈假单胞菌来检查体内宿主和细菌的相互作用。由于小鼠对牙龈假单胞菌的反应很幼稚，我们可以检查当不存在获得性免疫反应时的反应，并将其与通过牙龈假单胞菌预免疫激活适应性免疫反应的小鼠进行比较。通过流式细胞术，我们将详细分析FOXO1缺失在产生全身和局部适应性免疫反应中的影响。下面的颅骨也将进行组织学检查，以调查随后对破骨细胞形成和骨吸收的影响。实验小鼠(CD11cCre+/FOXO1L/L)和对照小鼠(CD11cCre-/FOXO1L/L)将牙龈假单胞菌应用于口腔内，以确定DCs中的FOXO1缺失是否调节宿主对牙龈假单胞菌的反应、破骨细胞的发生和牙周疾病的进展。使用相同的两个体内模型的实验将确定Akt是否对于预防DC的高炎症反应是必要的。这些实验将检测实验组(CD11cCre+/FOXO1L/L)和对照组(CD11cCre-/FOXO1L/L)小鼠。在目标3中，FOXO1-Akt轴调节选定的靶基因(IL-1、肿瘤坏死因子、IL-6和IL-12)表达的机制将在体外进行研究。